Identification of a novel AP-2 consensus DNA binding site.
Activator Protein (AP)-2 is a transcription factor that is required for mouse development. AP-2 activates expression of positive and negative growth regulators including erbB-2 and p21 WAF1/CIP1. Induction of p21 has been correlated with cell cycle and growth inhibition of human cancer cells. Because several endogenous AP-2 binding sites do not fit the known consensus sequences well, we sought to define AP-2's interaction with DNA more precisely. Using Cyclic Amplification and Selection of Targets (CAST'ing) of random oligonucleotide sequences and recombinant human AP-2 protein, we identified 17 novel AP-2 binding sites. Mobility shift assays showed significant AP-2 binding of the novel sites as compared to p21, erbB-2 and hMtIIa sites. Several sites that bound with high specificity and affinity did not fit known AP-2 consensus sequences. A sequence comparison based on several of the novel sequences yielded a putative consensus binding sequence of 5'-TAGAAAGNYCYNG-3'. These DNA binding sites may help identify novel targets of AP-2 and aid in further understanding AP-2 function.
Gee
M S
MS
Howard Hughes Medical Institute, Department of Medicine and Genetics, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Sarkisian
C J
CJ
el-Deiry
W S
WS
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D051379:Mice; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D004247:DNA